Bacteriolytic complex, method for producing said complex and strain for carrying out said method

Abstract

The inventions relate to the field of medicine, veterinary and biotechnology and may be used for production of a drug for medical and veterinary purposes. A bacteriolytic complex, produced by the bacterium Lysobacter sp. XL1, has been proposed, which contains bacteriolytic enzymes (muramidase, muramoylalanineamidase, endopeptidase, a bacteriolytic enzyme with a molecular weight of about 22 kDa), protease, polysaccharide, and ballast components. The method of production of the bacteriolytic complex includes cultivation of the strain-producer on a nutrient medium containing glucose, peptone, yeast extract or yeast autolysate, phosphate salts of sodium and potassium, magnesium sulfate, potassium chloride, iron sulfate, and water.

Description

TECHNICAL FIELD [0001]The inventions relate to the field of biotechnology, medicine, and veterinary, more precisely concerning a bacteriolytic complex, and may be used for production of a drug for medicine and veterinary.BACKGROUND[0002]It is known that bacteriolytic preparations are used as drugs in treatment for a number of diseases caused by pathogenic microflora, including that resistant to antibiotics.[0003]The preparation having bacteriolytic effect—lysozyme is known (Egorov N. S. Bases of the doctrine of antibiotics, 1979, “Vysshaya shkola”, p. 347–349). Lysozyme of egg protein is active against a narrow spectrum of microorganisms, limited mainly by micrococci. By specificity of action on microbial cell wall peptidoglycan, lysozyme is a muramidase, i.e., lyzes the bond between muramic acid and glucosamine with the formation of fragments with muramic acid on the reducing end.[0004]That is why lysozyme lyzes the narrow spectrum of gram-positive microorganisms. In particular, it...

AI Technical Summary

Benefits of technology

"The patent text describes a method for producing a complex of bacteriolytic enzymes using a strain of bacteria called Lysobacter sp. XL1. This method involves cultivating the bacteria on a nutrient medium containing specific components, and then precipitating the enzyme complex from the culture liquid. The resulting complex has a higher content of bacteriolytic enzymes and lower content of inactive proteins. The technical effects of this method include increased yield and productivity of the bacteriolytic enzymes, optimization of the composition of the end product, and acceleration of the production process. The price of the nutrient medium is lower, and the content of bacteriolytic enzymes in the culture liquid is raised. The method also reduces the amount of inactive components in the complex. Overall, this patent aims to develop a more efficient and cost-effective method for producing bacteriolytic enzymes."

Problems solved by technology

Besides, like most enzyme preparations, lyzozyme is not stable enough.

However, it is unstable and applied in laboratory practice only.

The disadvantage of this complex is that its bacteriolytic activity is not high enough due to the presence of ballast components and because nearly the half of the bacteriolytic enzymes are denatured.

Besides, the ballast components have no medicinal effect but may cause allergic reactions.

Both the content of bacteriolytic enzymes in culture liquid and the yield of finished preparation are not high enough at realization of this method.

The yield of end product is not high enough—about 20% of the lytic complex content in culture liquid.

The above method has a disadvantage that the protein-vitamin concentrate is currently not produced by domestic industry, because its production is ecologically unsafe; the use of expensive imported bactopeptone Difco (USA) as a medium component is unpractical, because it significantly increases the cost of the final product.

Besides, the use of ethanol (RU 1755581) and acetone (RU 1549227) for precipitation of the enzymes from culture liquid of the producer results in irreversible denaturing of about the half of bacteriolytic enzymes determining the therapeutic effect of the drug, which are present in the culture liquid (according to the results of bacteriolytic activity measurement and electrophoretic analysis) and, consequently, in a considerable decrease in the product yield (FIG. 1).

Finished lysoamidase preparations produced by the known methods contain a significant amount of minor components—ballast proteins (FIG. 1), which have no medicinal effect but may cause an allergic reaction.

The disadvantage of this strain-producer is that, due to the presence of minor components in the produced complex, bacteriolytic activity is insufficiently high.

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